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http://dx.doi.org/10.11626/KJEB.2021.39.4.415

Development of simultaneous detection method for living modified cotton varieties MON757, MON88702, COT67B, and GHB811  

Il Ryong Kim (Division of Ecological Safety, National Institute of Ecology(NIE))
Min-A Seol (Division of Ecological Safety, National Institute of Ecology(NIE))
A-Mi Yoon (Division of Ecological Safety, National Institute of Ecology(NIE))
Jung Ro Lee (Division of Ecological Safety, National Institute of Ecology(NIE))
Wonkyun Choi (Division of Ecological Safety, National Institute of Ecology(NIE))
Publication Information
Korean Journal of Environmental Biology / v.39, no.4, 2021 , pp. 415-422 More about this Journal
Abstract
Cotton is an important fiber crop, and its seeds are used as feed for dairy cattle. Crop biotechnology has been used to improve agronomic traits and quality in the agricultural industry. The frequent unintentional release of LM cotton into the environment in South Korea is attributed to the increased application of living modified (LM) cotton in food, feed, and processing industries. To identify and monitor the LM cotton, a method for detecting the approved LM cotton in South Korea is required. In this study, we developed a method for the simultaneous detection of four LM cotton varieties, MON757, MON88702, COT67B, and GHB811. The genetic information of each LM event was obtained from the European Commission-Joint Research Centre and Animal and Plant Quarantine Agency. We designed event-specific primers to develop a multiplex PCR method for LM cotton and confirmed the specific amplification. Using specificity assay, random reference material(RM) mixture analysis and limit of detection(LOD), we verified the accuracy and specificity of the multiplex PCR method. Our results demonstrate that the method enabled the detection of each event and validation of the specificity using other LM RMs. The efficiency of multiplex PCR was further verified using a random RM mixture. Based on the LOD, the method identified 25 ng of template DNA in a single reaction. In summary, we developed a multiplex PCR method for simultaneous detection of four LM cotton varieties, for possible application in LM volunteer analysis.
Keywords
living modified organisms; LMO; multiplex PCR; cotton;
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